Abstract
Human embryonic stem cells (hESC) are the subject of intense investigation for use in regenerative
medicine, in toxicity testing, and as models for the study of human development.
Automated cell sorting will enhance the isolation of homogenous pools of differentiated
hESCs both for basic studies and for therapeutic applications. Sorting could also be used to
deplete undifferentiated, potentially tumourigenic cells. However, hESCs are sensitive to single
cell disaggregation and recover poorly when plated at clonal density. Here we report a
method for successful semi-automated single cell sorting of hESCs. This method utilizes an
ES-specific promoter-transgene construct and automated FACS-based single cell sorting and
plating. Clonal recovery in physiologic oxygen (2%) was increased fourfold over room oxygen
(21%;
Get full access to this article
View all access options for this article.